新型复合水凝胶对H₂O₂诱导心肌细胞氧化应激损伤的保护作用  

Protective effect of novel composite hydrogels on H₂O₂-induced oxidative stress injury in cardiomyocytes

作　　者：王岳 马宁 卢嘉骏 王成瑶 陈琳渝 任宇晨 李景武[3] 孙红 WANG Yue;MA Ning;LU Jiajun;WANG Chengyao;CHEN Linyu;REN Yuchen;LI Jingwu;SUN Hong(Key Laboratory of Chronic Diseases,School of Basic Medical Sciences,North China University of Science and Technology,Tangshan 063210,China;Key Laboratory of Rehabilitation Engineering and Regenerative Medicine,School of Nursing and Rehabilitation,North China University of Science and Technology,Tangshan 063000,China;Key Laboratory of Molecular Oncology,Hebei Provincial People’s Hospital,Tangshan 063000,China)

机构地区：[1]华北理工大学基础医学院河北省慢性病重点实验室,河北唐山063210 [2]华北理工大学护理与康复学院河北省康复工程与再生医学重点实验室,河北唐山063000 [3]河北省唐山市人民医院河北省分子肿瘤学重点实验室,河北唐山063000

出　　处：《吉林大学学报(医学版)》2025年第2期352-359,共8页Journal of Jilin University:Medicine Edition

基　　金：国家自然科学基金项目(U20A20261);河北省科技厅自然科学基金项目(H2023209021,H2024209013);河北省教育厅大学生创新创业训练计划项目(202410081057);河北省唐山市科技局重点研发科技项目(22150201A)。

摘　　要：目的:探讨新型复合水凝胶对过氧化氢(H₂O₂)诱导心肌细胞氧化应激损伤的保护作用,并阐明其可能的作用机制。方法:小鼠皮下注射水凝胶100μL,1、14和28 d正常饲养后处死小鼠,取材,HE染色观察水凝胶的组织相容性。提取出生1 d SD大鼠的原代心肌细胞,建立心肌细胞氧化应激损伤模型。原代心肌细胞分为对照组、H₂O₂组和水凝胶(H₂O₂+Hydrogel)组。对照组原代心肌细胞正常培养,H₂O₂组原代心肌细胞采用200μmol·L-1 H₂O₂溶液处理24 h;H₂O₂+Hydrogel组原代心肌细胞采用1 g·L-1的水凝胶与200μmol·L-1 H₂O₂共孵育24 h。采用细胞计数试剂盒8(CCK-8)法检测各组心肌细胞活性,双氢乙啶(DHE)和2′,7′-二氯荧光素二乙酸酯(DCFH-DA)检测各组心肌细胞中活性氧(ROS)水平,鬼笔环肽荧光染色标记法检测各组心肌细胞中纤维形肌动蛋白(F-actin)表达情况,免疫荧光法检测各组心肌细胞中连接蛋白43(Cx43)和心肌肌钙蛋白T(cTnT)表达情况,TUNEL染色法检测各组心肌细胞凋亡率,Western blotting法检测各组心肌细胞中凋亡相关蛋白B细胞淋巴瘤2(Bcl-2)和Bcl-2相关X蛋白(Bax)的表达水平。结果:HE染色,水凝胶周围组织炎性细胞浸润较少,皮下埋植的炎症反应较小。与对照组比较,H₂O₂组心肌细胞活性明显降低(P<0.05),细胞中ROS水平明显升高(P<0.05),细胞中Cx43、cTnT和F-actin蛋白表达水平明显降低(P<0.001),心肌细胞凋亡率明显升高(P<0.01),心肌细胞中Bcl-2蛋白表达水平升高(P<0.001),Bax蛋白表达水平降低(P<0.01);与H₂O₂组比较,H₂O₂+Hydrogel组心肌细胞活性明显升高(P<0.05),细胞中ROS水平明显降低(P<0.01),细胞中cTnT、Cx43和F-actin蛋白表达水平明显升高(P<0.01),心肌细胞凋亡率明显降低(P<0.001),心肌细胞中Bcl-2蛋白表达水平降低(P<0.01),Bax蛋白表达水平升高(P<0.01)。结论:水凝胶能够通过清除环境中的ROS促进心肌细胞相关�Objective:To investigate the protective effect of a composite hydrogel against hydrogen peroxide(H₂O₂)-induced oxidative stress injury in the cardiomyocytes,and to clarify its possible mechanism.Methods:The mice were subcutaneously injected with 100μL of hydrogel.After normal feeding for 1,14,and 28 d,the mice were sacrificed.Tissue samples were collected and subjected to HE staining to observe the histocompatibity of the hydrogel.The primary cardiomyocytes isolated from 1-day-old SD rats were used to establish an oxidative stress injury model.The primary cardiomyocyties were divided into control,H2O2 and H2O2+Hydrogel groups.The primary cardiomyocytes in control group were cultured normally,the primary cardiomyocytes in H2O2 group were treated with 200μmol·L^(-1) H2O2 for 24 h,and the primary cardiomyocytes in H2O2+Hydrogel group were incubated with 1 g·L^(-1) composite hydrogel and 200μmol·L^(-1) H2O2 for 24 h.The viabilities of cardiomyocytes in various groups were assessed by cell counting kit-8(CCK-8)method.Dihydroethidium(DHE)and 2',7'-dichlorodihydrofluorescein diacetate(DCFH-DA)staining were used to assess the reactive oxygen species(ROS)levels in the cells.The expressions of filamentous actin(F-actin)in the cells in various groups were detected by phalloidin fluorescence staining;the expressions of connexin 43(Cx43)and cardiac troponin T(cTnT)proteins in the cardiomyocytes in various groups were detected by immunofluorescence method.The apoptotic rates of cardiomyocytes in various groups were assessed with TUNEL staining method.The expression levels of apoptosis-related proteins B-cell lymphoma-2(Bcl-2)and Bcl-2-associated X protein(Bax)in the cardiomyocytes in various groups were assessed by Western blotting method.Results:The HE staining results showed that the inflammatory cells around the implanted hydrogel were less infiltrated,and the inflammatory reaction of subcutaneous implantation was less.Compared with control group,the viability of cardiomyocytes in H2O2 group was significantly decr

关 键 词：水凝胶 心肌细胞 过氧化氢 氧化应激损伤 细胞凋亡 

分 类 号：R542.2[医药卫生—心血管疾病]